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    puma-antibody-ab9645.pdf

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Cell Biology Apoptosis Intracellular Bcl2 Family
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Anti-PUMA antibody (ab9645)

  • Datasheet
  • SDS
Submit a review Q&A (1)References (9)

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Western blot - Anti-PUMA antibody (ab9645)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PUMA antibody (ab9645)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PUMA antibody (ab9645)
  • Immunocytochemistry/ Immunofluorescence - Anti-PUMA antibody (ab9645)
  • Immunocytochemistry/ Immunofluorescence - Anti-PUMA antibody (ab9645)

Key features and details

  • Rabbit polyclonal to PUMA
  • Suitable for: WB, ICC/IF, IHC-P
  • Reacts with: Human
  • Isotype: IgG

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Secondary
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Goat Anti-Rabbit IgG H&L (HRP) (ab205718)

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Overview

  • Product name

    Anti-PUMA antibody
    See all PUMA primary antibodies
  • Description

    Rabbit polyclonal to PUMA
  • Host species

    Rabbit
  • Specificity

    Specific to PUMA alpha
  • Tested Applications & Species

    Application Species
    ICC/IF
    Human
    IHC-P
    Human
    WB
    Human
    See all applications and species data
  • Immunogen

    Synthetic peptide:

    ARARQEGSSPEPVEG

    , corresponding to amino acids 2-16 of Human PUMA. (Peptide available as ab9646).
    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • Positive control

    • WB: K562 lysate IHC-P: Human breast carcinoma sections ICC/IF: K562 cells
  • General notes


    Apoptosis is related to many diseases and development. The p53 tumor-suppressor protein induces apoptosis through transcriptional activation of several genes. A novel p53 inducible pro-apoptotic gene was identified recently and designated PUMA (for p53 upregulated modulator of apoptosis) and bbc3 (for Bcl-2 binding component 3) in human and mouse (1-3). PUMA/bbc3 is one of the pro-apoptotic Bcl-2 family members including Bax and Noxa, which are also transcriptional targets of p53. The PUMA gene encodes two BH3 domain-containing proteins termed PUMA-a and PUMA-b (1). PUMA proteins bind Bcl-2, localize to the mitochondria, and induce cytochrome c release and apoptosis in response to p53. PUMA may be a direct mediator of p53-induced apoptosis.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Storage buffer

    pH: 7.2
    Preservative: 0.02% Sodium azide
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Primary antibody notes

    Apoptosis is related to many diseases and development. The p53 tumor-suppressor protein induces apoptosis through transcriptional activation of several genes. A novel p53 inducible pro-apoptotic gene was identified recently and designated PUMA (for p53 upregulated modulator of apoptosis) and bbc3 (for Bcl-2 binding component 3) in human and mouse (1-3). PUMA/bbc3 is one of the pro-apoptotic Bcl-2 family members including Bax and Noxa, which are also transcriptional targets of p53. The PUMA gene encodes two BH3 domain-containing proteins termed PUMA-a and PUMA-b (1). PUMA proteins bind Bcl-2, localize to the mitochondria, and induce cytochrome c release and apoptosis in response to p53. PUMA may be a direct mediator of p53-induced apoptosis.
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Apoptosis
    • Intracellular
    • Bcl2 Family
    • Signal Transduction
    • Metabolism
    • Mitochondrial
    • Cancer
    • Invasion/microenvironment
    • Apoptosis
    • Bcl 2 family
    • Metabolism
    • Pathways and Processes
    • Mitochondrial Metabolism
    • Mitochondrial markers
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Apoptosis
    • Metabolism
    • Types of disease
    • Cancer
    • Cancer
    • Cell Death
    • Apoptosis
    • Apoptosis Markers
    • Bcl 2 family
    • Cancer
    • Cell Death
    • Apoptosis
    • Metabolism

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Positive Controls

    • K-562 whole cell lysate (ab7911)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab9645 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
ICC/IF
Human
IHC-P
Human
WB
Human
All applications
Mouse
Rat
Application Abreviews Notes
WB
Use a concentration of 2 - 4 µg/ml. Detects a band of approximately 23 kDa.
ICC/IF
Use a concentration of 20 µg/ml.
IHC-P
Use a concentration of 10 µg/ml.
Notes
WB
Use a concentration of 2 - 4 µg/ml. Detects a band of approximately 23 kDa.
ICC/IF
Use a concentration of 20 µg/ml.
IHC-P
Use a concentration of 10 µg/ml.

Target

  • Function

    Essential mediator of p53-dependent and p53-independent apoptosis.
  • Tissue specificity

    Ubiquitously expressed.
  • Sequence similarities

    Belongs to the Bcl-2 family.
  • Cellular localization

    Mitochondrion. Localized to the mitochondria in order to induce cytochrome c release.
  • Target information above from: UniProt accession Q9BXH1 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 27113 Human
    • Entrez Gene: 170770 Mouse
    • Entrez Gene: 317673 Rat
    • Omim: 605854 Human
    • SwissProt: Q9BXH1 Human
    • SwissProt: Q99ML1 Mouse
    • SwissProt: Q80ZG6 Rat
    • Unigene: 467020 Human
    • Unigene: 7660 Mouse
    • Unigene: 25176 Rat
    see all
  • Alternative names

    • BBC 3 antibody
    • Bbc3 antibody
    • BBC3_HUMAN antibody
    • BCL 2 binding component 3 antibody
    • Bcl-2-binding component 3 antibody
    • BCL2 binding component 3 antibody
    • JFY 1 antibody
    • JFY-1 antibody
    • JFY1 antibody
    • p53 up regulated modulator of apoptosis antibody
    • p53 up-regulated modulator of apoptosis antibody
    • p53 Upregulated Modulator of Apoptosis antibody
    • PUMA alpha antibody
    • PUMA/JFY1 antibody
    see all

Images

  • Western blot - Anti-PUMA antibody (ab9645)
    Western blot - Anti-PUMA antibody (ab9645)
    Anti-PUMA antibody (ab9645) at 2 µg/ml + K562 cell lysates at 15 µg

    Secondary
    Goat anti-rabbit IgG HRP conjugate at 1/10000 dilution


    Primary incubation: 1 hour at room temperature in 5% NFDM/TBST. 

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PUMA antibody (ab9645)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PUMA antibody (ab9645)

    Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue labeling PUMA with ab9645 at 10 μg/mL. Sections were fixed with formaldehyde and blocked with 10% serum for 1 hour at room temperature. Antigen retrieval was by heat mediation with a citrate buffer (pH 6.0). Samples were incubated with primary antibody overnight at 4˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PUMA antibody (ab9645)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PUMA antibody (ab9645)

    Immunohistochemical analysis of paraffin-embedded human breast tissue labeling PUMA with ab9645 at 2.5 μg/mL. Sections were fixed with formaldehyde and blocked with 10% serum for 1 hour at room temperature. Antigen retrieval was by heat mediation with a citrate buffer (pH 6.0). Samples were incubated with primary antibody overnight at 4˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

  • Immunocytochemistry/ Immunofluorescence - Anti-PUMA antibody (ab9645)
    Immunocytochemistry/ Immunofluorescence - Anti-PUMA antibody (ab9645)

    Immunofluorescent analysis of 4% paraformaldehydefixed K562 cells labeling PUMA with ab9645 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red) and DAPI staining (blue).

  • Immunocytochemistry/ Immunofluorescence - Anti-PUMA antibody (ab9645)
    Immunocytochemistry/ Immunofluorescence - Anti-PUMA antibody (ab9645)

    Immunofluorescent analysis of 4% paraformaldehydefixed K562 cells labeling PUMA with ab9645 at 10 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red). Image showing cytosol staining on K562 cells.

Protocols

  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (9)

Publishing research using ab9645? Please let us know so that we can cite the reference in this datasheet.

ab9645 has been referenced in 9 publications.

  • Mensink M  et al. Anti-apoptotic A1 is not essential for lymphoma development in Eµ-Myc mice but helps sustain transplanted Eµ-Myc tumour cells. Cell Death Differ 25:795-806 (2018). WB ; Mouse . PubMed: 29339775
  • Palomo GM  et al. Silencing of frataxin gene expression triggers p53-dependent apoptosis in human neuron-like cells. Hum Mol Genet 20:2807-22 (2011). PubMed: 21531789
  • Höglund A  et al. Myc sensitizes p53-deficient cancer cells to the DNA-damaging effects of the DNA methyltransferase inhibitor decitabine. Blood 113:4281-8 (2009). WB, IF ; Human, Mouse . PubMed: 19179467
  • Lim LY  et al. Mutant p53 mediates survival of breast cancer cells. Br J Cancer 101:1606-12 (2009). WB ; Human . PubMed: 19773755
  • Harford TJ  et al. Increased expression of the pro-apoptotic Bcl2 family member PUMA and apoptosis by the muscle regulatory transcription factor MyoD in response to a variety of stimuli. Apoptosis : (2009). WB ; Mouse . PubMed: 19943111
  • Leong CO  et al. The p63/p73 network mediates chemosensitivity to cisplatin in a biologically defined subset of primary breast cancers. J Clin Invest 117:1370-80 (2007). PubMed: 17446929
  • Bouchard C  et al. FoxO transcription factors suppress Myc-driven lymphomagenesis via direct activation of Arf. Genes Dev 21:2775-87 (2007). PubMed: 17974917
  • Biswas SC  et al. Puma and p53 play required roles in death evoked in a cellular model of Parkinson disease. Neurochem Res 30:839-45 (2005). PubMed: 16187218
  • Stankovic T  et al. Microarray analysis reveals that TP53- and ATM-mutant B-CLLs share a defect in activating proapoptotic responses after DNA damage but are distinguished by major differences in activating prosurvival responses. Blood 103:291-300 (2004). PubMed: 12958068

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

Question

Thank you for getting back to me. We understand that you cannot give us a sample of an antibody to test. We had been using ab9643, but in light of our and other labs problems with the antibody, we are looking for a new PUMA antibody, specifically ab1074 or ab9645). Since you can't send us a sample, can you send us references of labs that have used these antibodies?

Read More

Abcam community

Verified customer

Asked on Oct 31 2003

Answer

I'm sorry, but I can not give out that type of information. We strongly encourage our customers to give us feedback on our antibodies and when we do receive such feedback, we post those reviews on the appropriate datasheets. Unfortunately in the case of ab9645 we do not have any references or reviews.

Read More

Abcam Scientific Support

Answered on Oct 31 2003

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

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